Freddy Benning

10/💰 This study was made possible by funding from the NIH, CIHR, CFI, NSERC, NFRF, HHWF, @moorefound.bsky.social @simonsfoundation.org @snsf.ch 🇺🇸🇨🇦🇨🇭

9/ 📣 This was a wonderful collaboration with the daCosta Lab at @uottawa.ca.

A big shout-out to my 🦋-less co-firsts Melody Nguyen and Tristan Bell and to all co-authors. @lukechao.bsky.social

Thanks to the HMS EM facility, HMS MIcRoN, and the MIT.nano cryo-EM facility for their support.

8/🎯 Our observations reflect a lack of correlation between respiratory function and cristae attachment.

Instead, we observe other morphological cristae defects associated with compromised respiration.

By identifying functional key residues, this work paves the way for future mechanistic studies.

7/🔬 Using TEM, cryo-ET and fluorescence microscopy, we describe a phenotype that is specific to a mutant of these four ASR-identified residues, which features box-like shaped cristae and longer average cristae lengths.

6/🔬 Several TEM studies have shown that Mic60 and Mitofilin deletions result in detached cristae. We observe the same phenotype for our knockout variants. ✅

🤯 Surprisingly, Mitofilin ancestors which rescue respiratory growth like wildtype have knockout-like detached cristae.

5/🎯 Comparing reconstructed ancestors, we identified four residues in the Mitofilin domain that are sufficient for rescuing respiration.

4/🧫 We found that common Mitofilin ancestors as far back as the putative last opisthokont common ancestor #LOCA rescues respiration in yeast. ✅

3/❓How did Mitofilin-supported respiration diverge between yeast 🍺 and humans 👤?

💻🦖🧪 We used ancestral sequence reconstruction #ASR to reconstruct ancestral Mitofilin sequences and resurrected them for in vivo characterization.

2/ 🧫 Using respiratory growth assays, we found that swapping the yeast Mitofilin domain with that of another yeast rescued respiration. ✅

Replacing it with human or bacterial homologues at the endogenous locus did not rescue respiration. 💀

❗This is in contrast to previous plasmid-based work.

1/ ❓Mic60, a core component of the #MICOS complex in the #mitochondrial inner membrane, is crucial for the organization of #cristae and their functions. It's unclear, how Mic60's #Mitofilin domain encodes cristae-supported functions like cellular respiration.

Ancestral sequence reconstruction of the Mic60 Mitofilin domain reveals residues supporting respiration in yeast In eukaryotes, cellular respiration takes place in the cristae of mitochondria. The mitochondrial inner membrane protein Mic60, a core component of the mitochondrial contact site and cristae organizi...

Out in print! Using ancestral sequence reconstruction 🦖, in vivo characterization, and 🔬 analysis, we identified key residues in the mitochondrial protein #Mic60 that affect #respiration in #yeast.

Since the preprint, we added some cool findings. 🧵⬇️

onlinelibrary.wiley.com/doi/10.1002/...

I’m thrilled to share that I will join @stowersinstitute.bsky.social this fall to explore the evolution and design of bioenergetic membranes in bacteria.

We’re proud to welcome two bold scientific minds to the Institute! Arvind Pillai, PhD, joins as Assistant Investigator exploring protein #evolution and design. Friederike Benning, PhD, joins as Stowers Fellow studying membrane complexity in #bacteria.

Read more: bit.ly/3SCZQiM

Congrats Chen and colleagues (with shoutout to @myfry.bsky.social for connecting our labs)! JCI - SARM1 loss protects retinal ganglion cells in a mouse model of Autosomal Dominant Optic Atrophy www.jci.org/articles/vie...

🚨👉 Please check our recent work on bacterial cell division. In situ Cryo-ET reveals the cellular function of the penicillin binding protein 1b supported by AFM, live-cell imaging, in silico AlphaFold proteome screen and TIRFM. Hope you enjoy the read! #teamtomo #cryo-ET ❄️🔬🐎 big thanks to the team!

Interested in learning more about genome editing tool dev & their application to treat human diseases? 🧬 🏥
We're hiring graduating seniors who are seeking ~2 years of post-bac research experience.
If you're interested in gaining experience in next gen medicines, please reach out and apply!
Link 👇

Great summary of our recent preprint by Barbora from @prelights.bsky.social. #mitochondria #cristae #respiration #asr

A big shout-out to my incredible team mates Simon Jenni, Coby Garcia, Melody Nguyen, Xuewu Zhang and @lukechao.bsky.social

Helical reconstruction of VP39 reveals principles for baculovirus nucleocapsid assembly - Nature Com... Baculoviruses are used as insecticides, gene therapy tools, and protein production vectors. Here, the authors report the structure of the Autographa californica MNPV nucleocapsid and its fold conserva...

How do baculoviruses protect their genome?

Delighted to present our findings in print!

We use cryo-EM to determine the structure of a baculoviral nucleocapsid. We show how the tiny capsid protein VP39 assembles into a giant nucleocapsid.❄️🔬

www.nature.com/articles/s41...